[1]王延映,彭麟,余祖功.高效液相色谱法测定饲料中甲硝唑、地美硝唑和异丙硝唑[J].南京农业大学学报,2014,37(5):123-127.[doi:10.7685/j.issn.1000-2030.2014.05.020]
 WANG Yanying,PENG Lin,YU Zugong.Determination of metronidazole,dimetridazole and ipronidazole in feeds by HPLC[J].Journal of Nanjing Agricultural University,2014,37(5):123-127.[doi:10.7685/j.issn.1000-2030.2014.05.020]
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高效液相色谱法测定饲料中甲硝唑、地美硝唑和异丙硝唑()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
37卷
期数:
2014年5期
页码:
123-127
栏目:
出版日期:
2014-09-25

文章信息/Info

Title:
Determination of metronidazole,dimetridazole and ipronidazole in feeds by HPLC
作者:
王延映 彭麟 余祖功
南京农业大学动物医学院, 江苏 南京 210095
Author(s):
WANG Yanying PENG Lin YU Zugong
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
关键词:
甲硝唑地美硝唑异丙硝唑饲料高效液相色谱法
Keywords:
metronidazoledimetridazoleipronidazolefeedsHPLC
分类号:
S859.7
DOI:
10.7685/j.issn.1000-2030.2014.05.020
摘要:
为建立能同时检测饲料中甲硝唑、地美硝唑和异丙硝唑的简便、准确的高效液相色谱方法,样品用乙酸乙酯提取、MCX固相萃取柱净化、ZORBAX SB-C18柱(4.6 mm×250 mm,5 μm)分离,以甲醇-水为流动相,梯度洗脱,320 nm紫外检测,外标法定量。结果表明:此条件下,3种药物分离良好,标准曲线回归系数为0.999 7~0.999 8,线性范围0.02~10 μg·mL-1;平均回收率75.3%~106.3%,批内变异系数1.3%~12.4%,批间变异系数2.4%~9.7%,甲硝唑、地美硝唑和异丙硝唑检测限均为0.01 mg·kg-1,定量限为0.02 mg·kg-1。结论:该法操作简单、结果准确,可用于测定饲料中甲硝唑、地美硝唑和异丙硝唑的含量。
Abstract:
The purpose of this study was to develop a simple and effective method that optimized the high-performance liquid chromatography(HPLC)method for determination of metronidazole, dimetridazole and ipronidazole in feeds by investigating the extraction and clean-up conditions. Samples were weighed into 50 mL polypropylene centrifuge tubes and extracted by mechanical shaking for 30 min with 15 mL ethyl acetate. After extraction, the content was centrifuged at 6 000 r·min-1 for 10 min and the extract was transferred to a new polypropylene centrifuge tube. The unsolved feed matrix was extracted again by adding 15 mL ethyl acetate and the extract was collected. Both obtained ethyl acetate extractions were combined and evaporated just to dryness with a pressure blowing concentrator(water bath at 35 ℃). The residue was dissolved in 300 μL ethyl acetate and 4 mL hexane and the solution was transferred to a 10 mL polypropylene centrifuge tube first. Then 1.5 mL phosphoric acid solution(0.1 mol·L-1)was added into the tube. After vortexing(1 min)and centrifuging(10 min, 8 000 r·min-1), the lower aqueous phase was collected in a 5 mL polypropylene centrifuge tube. The supernatant organic phase was extracted again by adding 1.5 mL phosphoric acid solution(0.1 mol·L-1)and both the obtained lower aqueous phases were combined. The obtained solution was purified using mixed-mode cation exchange(MCX)solid-phase extraction(SPE)column and filtered using a 0.22 μm filter. 10 μL were injected by the auto sampler and separated with UV detector 320 nm pumping at a flow rate of 0.8 mL·min-1. The liquid chromatographic separation of the extracts was performed on a C18 bonded silica column applying a gradient of elution with water(A)and methanol(B). The total run time was 20 min. Results showed that the limit of detection of metronidazole, dimetridazole and ipronidazole was 0.01 mg·kg-1, and the quantitation limit was 0.02 mg·kg-1. The recoveries of the metronidazole, dimetridazole and ipronidazole for feeds samples spiked with 0.02, 0.05, 0.5, 5 mg·kg-1 were in the range of 75.3%-106.3%. The linearity was satisfactory with a correlation coefficient of 0.999 7-0.999 8 at concentrations ranging from 0.02 to 10 μg·mL-1. The intra-day precision(n=5)for nitroimidazoles in feeds spiked at 0.02, 0.05, 0.5, 5 mg·kg-1 were in the range of 1.3%-12.4% and the inter-day precision for 3 days(n=5)was 2.4%-9.7%. Conclusion:This method was characterised simplicity of operator with accurate results, and thus could be used to determine the contents of metronidazole, dimetridazole and ipronidazole in feeds.

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备注/Memo

备注/Memo:
收稿日期:2014-3-29。
基金项目:农业行业标准制定与修订(农产品质量安全)项目(2013)
作者简介:王延映,硕士研究生。
通讯作者:余祖功,副教授,主要从事新兽药药理研究及兽药新制剂的研发,E-mail:yuzugong@njau.edu.cn
更新日期/Last Update: 1900-01-01